title: do25112
description: Protocols: Nuclei was isolated from frozen livers from young C57BL/6 mice (3 month old) using the protocol described in Krishnaswami et al. 2016 (Nature Protocols, 11: 499-524). Nuclei was purified using Iodixanol gradient (Optiprep, D1556, Sigma Aldrich Chemie). Nuclei were stained with Hoechst and FACS sorted according to their ploidy level diploid (2n) and tetraploid (4n) in 384-well plates. cDNA were made on 384-well plates using single-nucleus-RNA-seq2 protocol based on a modified version of SMART-Seq v4 Ultra Low input RNA (Takara). ERCC spike-in RNA (Ambion) (0.25 µL diluted at 1:100,000 and 1:300,000) was added to the RT-Mix (5µL final reaction). Sequencing libraries were prepared using Illumina Nextera XT kit (Illumina, #FC-131-1096) and the combination of 384 Combinatorial Dual Indexes (Illumina- Set A to D , #FC-131-2001 to FC-131-2004) following the manufacturer instructions but miniaturizing the reactions 10x using a liquid handling robot (Mosquito HV).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2021-05-07T00:26:49Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2020-07-20T14:59:00Z
attributes.tag: External Id
attributes.value: SAMEA7073784
attributes.tag: INSDC center name
attributes.value: Helmholtz Pioneer Campus
attributes.tag: INSDC first public
attributes.value: 2021-05-07T00:26:49Z
attributes.tag: INSDC last update
attributes.value: 2020-07-20T14:59:00Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-93331595253410:do25112
attributes.tag: age
attributes.value: 3
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: individual
attributes.value: 16/036335 (SNI-13)
attributes.tag: organism part
attributes.value: liver
attributes.tag: ploidy
attributes.value: 2n
attributes.tag: post analysis well quality
attributes.value: fail
attributes.tag: sample name
attributes.value: E-MTAB-93331595253410:do25112
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: male
attributes.tag: strain
attributes.value: C57Bl6/J